Field of the Invention
The present invention relates to natural beta-cryptoxanthin concentrates of high purity and a process for its preparation. More particularly, the present invention provides beta-cryptoxanthin concentrates containing about 10-80% by weight total xanthophylls (total carotenoids) of which the trans-beta-cryptoxanthin content is about 75-98% by weight and the remaining including zeaxanthin, trans-capsanthin, beta-carotene and trace amounts of other carotenoids. The concentrates are particularly useful as dietary supplements for nutrition and health promoting benefits.
The invention also provides a process for the preparation of the beta-cryptoxanthin concentrate from plant oleoresin, especially from Capsicum oleoresin. The process includes the steps of admixing the oleoresin with alcohol solvents, saponifying the xanthophyll esters, washing and purifying by eluting the crude xanthophyll viscous concentrate on a silica gel column and purifying further by washings to obtain high purity trans-beta cryptoxanthin enriched concentrate crystals.
Background
Carotenoids represent one of the most widespread groups of naturally occurring fat-soluble pigments imparting yellow, red and orange color in plants as well as in animals. These absorb light in the 400-500 nm region of the visible spectrum and have a common chemical feature, a poly-isoprenoid structure, a long conjugated chain of double bonds in the central portion of the molecule and near symmetry around the central double bond. The basic structure can be modified in a number of ways such as by cyclization of the end groups and by introduction of oxygen functions (O—H, C═O) to yield a large family of more than 600 compounds, exclusive of cis- and trans-isomers. Mammalian species do not synthesize carotenoids, and therefore they have to be obtained from dietary sources such as fruits and vegetables and/or dietary supplements.
Carotenoids are classified into hydrocarbon carotenoids, with lycopene and beta-carotene being the important members, and oxycarotenoids (xanthophylls), to which belongs mono-hydroxylated beta-cryptoxanthin while lutein, zeaxanthin and astaxanthin are dihydroxylated. The biosynthetic pathway of enzymatic hydroxylation of symmetrical beta-carotene (beta, beta-carotene) leads to the formation of beta-cryptoxanthin (beta,beta-caroten-3-ol) whereas the same reaction starting from asymmetric alpha-carotene (beta, epsilon-carotene) gives rise to two reaction products, namely: alpha-cryptoxanthin (beta, epsilon-caroten-3′-ol) and zeinoxanthin (beta, epsilon-caroten-3-ol). Often, because of the spectral structural similarities between alpha-cryptoxanthin and zeinoxanthin, their identification is difficult and misleading unless chemical reactions are carried out such as methylation or base catalyzed isomerization. Further, the chemical structures of these (see FIG. 1) are also given wrongly in many publications.
Among the 20 carotenoids detected in mammalian plasma and tissues, beta-cryptoxanthin is one of the major carotenoids detected along with lutein, zeaxanthin, beta-carotene and lycopene, together accounting for nearly 90% of the carotenoids. (J. G. Bieri, E. D. Brown and J. C. Smith, Determination of individual carotenoids in human plasma by HPLC, J. Liq. Chromatogr. 8, 473-484, 1985). Beta-cryptoxanthin, a provitamin A, plays an important role in the diet, finally converting in the human body into an active form of vitamin A (retinol), a nutrient important for vision, immune function, and skin and bone health. Beta-cryptoxanthin has about one half the vitamin A activity of the major vitamin precursor, beta-carotene. In addition, beta-cryptoxanthin acts as an antioxidant in the body. Wingerath et. al. (1995) studied the uptake of beta-cryptoxanthin after ingestion of tangerine juice concentrate rich in beta-cryptoxanthin esters. Beta-cryptoxanthin in substantial amounts was detected both in human chylomicrons and in serum (T. Wingerath, W. Stahl and H. Sies, Beta-cryptoxanthin selectively increases in human chylomicrons upon ingestion of tangerine concentrate rich in beta-cryptoxanthin, Arch. Biochem. and Biophys., 324, 385-390, 1995). The bioavailability of the carotenoids from paprika oleoresin has shown the presence of beta-carotene and beta-cryptoxanthin in higher amounts in chylomicrons compared to zeaxanthin among the volunteers (A. Perez-Galvez, H. D. Martin, H. Sies and W. Stahl, Incorporation of carotenoids from paprika oleoresin into human chylomicrons, J. Nutrition, 89, 787-793, 2003). Burri et al. (2011) have reported that beta-cryptoxanthin bioavailability seems to be 7-fold greater than beta-carotene under similar conditions. Therefore, beta-cryptoxanthin can likely be a valuable and potential source of vitamin A, which needs further study and confirmation (B. J. Burri, S. Jasmine, T. Chang and T. R. Neidlinger, Beta-cryptoxanthin- and alpha-carotene-rich foods have greater apparent bioavailability than beta-carotene-rich foods in Western diets, Brit. J. Nutrition, 105, 212-219, 2011).
Unlike other carotenoids, beta-cryptoxanthin is not found in most fruits or vegetables but is present in certain specific foods such as capsicum species, citrus fruits, mango, papaya, and pumpkin in small amounts, e.g., at about 10-20 mg/100 g in these fruits and vegetables. Mostly, beta-cryptoxanthin is present in an ester form in paprika and mandarin fruits. Breithaupt and Bamedi (2001) have analyzed a large number of fruits and vegetables and reported the beta-cryptoxanthin ester concentration levels. The highest ester concentrations were found in red chilies (17.1 mg/100 g), tangerine and oranges (Carotenoid esters in vegetables and fruits: A screening with emphasis on beta-cryptoxanthin esters, J. Agric. 49, 2064-2070, 2001). Later, Breithaupt et al., in a randomized, single-blind crossover study using a single dose of esterified or non-esterified beta-cryptoxanthin in equal amounts found no difference in the resulting plasma response among 12 volunteers suggesting a comparable bioavailability. (D. E. Breithaupt, P. Weller, M. Wolters and A. Hahn, Plasma response to a single dose of dietary Beta-cryptoxanthin ester from papaya, Carica papaya, or non-esterified beta-cryptoxanthin in adult human subjects: a comparative study, Brit. J. Nutr. 90, 795-801, 2003). Takayanagi and Mukai (2009) developed bioavailable composition of beta-cryptoxanthin derived from citrus unshiu Marc by using an enzyme process and in combination with dietary fiber (U.S. Pat. Appl. Pub. No. 2009/0258111, Highly bioavailable oral administration composition of cryptoxanthin).
In most of the Western countries and Japan, the dietary source of beta-cryptoxanthin comes from citrus fruits and their products and consequently the plasma beta-cryptoxanthin levels can be considered as a good index of the amount of fruit consumption. Similarly, in fruits such as papaya widely consumed in many tropical countries (e.g., Latin America), a high correlation of plasma beta-cryptoxanthin has been reported (M. S. Irwig, A. El-Sohemy, A. Baylin, N. Rifai and H. Campos, Frequent intake of tropical fruits that are rich in beta-cryptoxanthin is associated with higher plasma beta-cryptoxanthin concentrations in Costa Rican adolescents, J. Nutr. 132, 3161-3167, 2002).
The water soluble extract of marine algae extract, specifically Sargassum horneri showed an anabolic effect on bone calcification in the femoral-metaphysical tissue of young and old rats in vivo and in vitro, suggesting its role in the prevention of osteoporosis (Yamaguchi et al., Effect of marine algae extract on bone calcification in the femoral-metaphysical tissues of rats: Anabolic effect of Sargassum horneri, J. Health Sci., 47, 533-538, 2001; Uchiyama and Yamaguchi, Anabolic effect of marine alga Sargassum horneri, Effect on bone components in the femoral-diaphyseal and -metaphyseal tissues of young and old rats in vivo, J. Health Sci. 48, 325-330, 2002). Later, in a study of the effects of the various carotenoids, beta-cryptoxanthin showed a significant increase in calcium content and alkaline phosphatase activity in the femoral-diaphyseal (cortical bone) and femoral-metaphyseal (trabecular bone) tissues, suggesting that beta-cryptoxanthin possesses a unique anabolic effect on bone calcification in vitro (Yamaguchi and Uchiyama, Effect of carotenoid on calcium content and alkaline phosphatase activity in rat femoral tissues in vitro: The unique anabolic effect of beta-cryptoxanthin, Biol. Pharma. Bull. 26, 1188-1191, 2003). In another study, the DNA content in bone tissues was found to increase significantly and showed inhibitory effect on bone-resorbing factors-induced bone resorption in rat bone tissues in vitro (Uchiyama et al., Anabolic effect of beta-cryptoxanthin on bone components in the femoral tissues of aged rats in vivo and in vitro, J. Health Sci. 50, 491-496, 2004; Yamaguchi and Uchiyama, Beta-cryptoxanthin stimulates bone formation and inhibits bone resorption in tissue culture in vitro, Mol. Cell. Biochem. 258, 137-144, 2004).
Thus, beta-cryptoxanthin has a potential role and effect in maintaining bone health and preventing osteoporosis. The various studies carried out by Yamaguchi et al. (2004, 2005, 2006 and 2008), have shown that regular daily intake of Satsuma mandarin juice (Citrus unshiu) and/or supplemented with beta-cryptoxanthin (3-6 mg or more/day) has beneficial effects such as preventive effect on bone loss over age, stimulatory effect on bone formation and an inhibitory effect on bone re-absorption in normal and healthy individuals and in menopausal women (Prolonged intake of juice, Citrus unshiu, reinforced with beta-cryptoxanthin has an effect on circulating bone biomarkers in normal individuals, J. Health Sci., 50, 619-624, 2004; Relationship between serum beta-cryptoxanthin and circulating bone metabolic markers in healthy individuals with the intake of juice (Citrus unshiu) containing beta-cryptoxanthin. J. Health Sci., 51, 738-743, 2005; Effect of beta-cryptoxanthin on circulating bone biomarkers intake of juice (Citrus unshiu) supplemented with beta-cryptoxanthin has effect in menopausal women, J. Health Sci., 52, 758-768, 2006; Beta-Cryptoxanthin and bone metabolism: The prevention role in osteoporosis, J. Health Sci., 54, 356-369, 2008). Uchiyama and Yamaguchi (2005 & 2006) reported that oral administration of beta-cryptoxanthin isolated from Satsuma mandarin had a preventive effect in bone loss in streptozotocin (diabetic) and ovariectomized rats in vivo studies (Oral administration of beta-cryptoxanthin prevents bone loss in streptozotocin rats in vivo, Biol. Pharm. Bull. 28, 1766-1769, 2005; Oral administration of beta-cryptoxanthin prevents bone loss in ovariectomized rats, Int. J. Mol. Med. 17, 15-20, 2006). EP Application No. 058060229, Publn. 2007/35 (published as EP Publn. EP1825858) refers to a composition comprising beta-cryptoxanthin and zinc for promoting osteogenesis, increasing bone mineral content, thereby preventing bone diseases such as osteoarthritis. (M. Yamaguchi, Composition for promoting osteogenesis and increasing bone mineral content). U.S. Pat. No. 8,148,431 B2, Apr. 3, 2012, confirm that beta-cryptoxanthin has an osteogenesis promoting effect, a bone-resorption inhibiting effect and therapeutic effect on bone diseases (M. Yamaguchi, Osteogenesis promoter containing beta-cryptoxanthin as the active ingredient).
Generally, the different bone and joint disorders such as osteoporosis, osteoarthritis and rheumatoid arthritis are common among the elderly people and cause a major health problem resulting in bone fracture. With ageing there is a decrease in bone mass and an increase in bone resorption due to various dietary reasons. In recent publications, Yamaguchi has reviewed the recent advances concerning the role of beta-cryptoxanthin in the regulation of bone homeostasis and in the prevention of osteoporosis, especially the cellular and molecular mechanisms by which beta-cryptoxanthin stimulates osteoblastic bone formation and inhibits osteoclastic bone resorption (Beta-cryptoxanthin and bone metabolism: The preventive role in osteoporosis, J. Health Sci., 54, 356-369, 2008; Role of carotenoid beta-cryptoxanthin in bone homeostasis, J. Biomed. Sci., 19, 1-13, 2012).
High levels of dietary intake of beta-cryptoxanthin were found to be associated with reduced risk of lung cancer among the smoking population, thereby suggesting the xanthophylls as a chemo-preventive agent for lung cancer (Yuan et al., Dietary cryptoxanthin and reduced risk of lung cancer: the Singapore Chinese health study, Cancer Epidemiol. Biomarkers Prev. 12, 890-898, 2003). Craft et al. (2004) found beta-cryptoxanthin in the frontal cortex of brain which is considered to be associated with Alzheimer's disease, however explained no exact role for beta-cryptoxanthin (N. E. Craft, T. B. Haitema, K. M. Garnett, K. A. Fitch and C. K. Dorey, Carotenoids, tocopherol and retinol concentrations in elderly human brain, J. Nutr. Health Ageing, 8, 156-162, 2004). A high plasma level of beta-cryptoxanthin has been linked to a protective effect against rheumatoid arthritis. Pattison et al. (2005) attributed the incidence of inflammatory arthritis among 88 subjects to the low level of dietary intake of beta-cryptoxanthin (D. J. Pattison, D. P. Symmons, M. Lunt, A. Welch, S. A. Bingham, N. E. Day and A. J. Silman, Dietary beta-cryptoxanthin and inflammatory polyarthritis: results from a population based prospective study, Am. J. Clin. Nutri. 82, 451-455, 2005). U.S. Pat. Appl. Pub. No. 2008/0070980 describes a method of use of beta-cryptoxanthin and its esters in the manufacture of a composition for providing increased protein formation and/or prevention of loss of proteins in human and animals, resulting in enhanced performance in sports and workout activities (E. Anne, G. Resina, W. Karin and W. Adrian, Use of beta-cryptoxanthin, Mar. 20, 2008). In a recent U.S. patent application entitled “Method of improving cardiovascular health,” there is a finding that a nutritional supplement containing purified beta-cryptoxanthin (0.1 to 20 mg/day) is effective in lowering high blood pressure and also in maintaining a healthy blood pressure and cardiovascular health. However, the beta-cryptoxanthin used is purified by analytical HPLC from a mixture of alpha-cryptoxanthin, anhydroluteins, zeaxanthin, and other impurities (U.S. Pat. Appl. Pub. No. 2012/0053247, Publn. 1 Mar. 2012, H. Showalter, Z. Defretas and L. Mortensen).
In view of the increasing research interest in the various health benefits of beta-cryptoxanthin, there have been several approaches to commercially produce this carotenoid (1) from natural sources as extracts rich in beta-cryptoxanthin, (2) by biotechnology routes and (3) by total- and semi-synthesis. The various clinical studies have used either synthetic beta-cryptoxanthin or natural fruit extracts rich in beta-cryptoxanthin.
Natural Source Extracts
Yamaguchi (2006) referred to a method for separating beta-cryptoxanthin from Satsuma orange by extracting the pigment using hydrolyzation followed by silica gel column chromatography. The beta-cryptoxanthin fraction was further purified by octadecyl silicate silica to obtain 95% beta-cryptoxanthin (M. Yamaguchi, Osteogenesis promoter containing beta-cryptoxanthin as an active ingredient, U.S. Pat. Appl. Pub. No. 2006/0106115, published 18 May 2006). Takahashi and Inada (2007) have prepared Persimmon extract from pulp/juice and skin by solvent extraction and hydrolysis to liberate beta-cryptoxanthin (free). The extracts prepared from pulp and skin showed 1 mg/100 g and 8 mg/100 g beta-cryptoxanthin, respectively, and useful applications in functional foods (H. Takahashi and Y. Inada, U.S. Pat. Appl. Pub. No. 2007/0116818 A, published 24 May 2007, Extract containing beta-cryptoxanthin from Persimmon fruit). Shirakura et. al (2008) and Takayanagi and Mukai (2008) have developed commercial processes for enzyme treated Satsuma mandarin (EPSM) and emulsified mandarin extract (EME) containing 0.2 and 0.05% beta-cryptoxanthin, respectively. They reported that the extracts possess reduction of visceral fat and plasma glucose in a human comparative trial designed as placebo-controlled double blind study (Y. S. Hirakura, K. Takayanagi and K. Mukai, Reducing effect of beta-cryptoxanthin extracted from Satsuma mandarin on human body fat, Abstract, page 161; K. Takayanang and K. Mukai, Abstract: Beta-cryptoxanthin and Satsuma mandarin: Industrial production and health promoting benefits, page, 73, Carotenoid Science, 12 Jun. 2008, Abstracts of the papers presented at the 15th International Symposium on Carotenoids, Okinawa, Japan, 22-27 Jun. 2008).
Biotechnological Production
Serrato-Joya et al. (2006) have generated beta-cryptoxanthin in a laboratory scale batch production using Flavobacterium lutescens ITC B 008 (O. Serrato-Joya, H. Jimenez-Islas, E. Botello-Alvarez, R. Nicomartinez, and J. L. Navarrete-Bolans, Process of beta-cryptoxanthin, a provitamin A precursor by Flavobacterium Lutescens, J. Food Sci. 71, E314-E319, 2006). Louie and Fuerst (2008) disclosed a method for preparing beta-cryptoxanthin from a microorganism transformed with the beta-carotene hydroxylase gene from Arabidopsis thaliana by culturing the transformant and recovering beta-cryptoxanthin (U.S. Pat. Appl. Pub. No. 2008/0124755, published 29 May 2008, Biosynthesis of beta-cryptoxanthin in microbial hosts using Arabidopsis thaliana beta-carotene hydroxylase gene). Again, Louie and Fuerst (2009) disclosed a method of beta-cryptoxanthin production by the use of lycopene beta-monocyclase and converting lycopene to beta-cryptoxanthin through gamma-carotene and 3-hydroxy-gamma-carotene (M. Y. Louie and E. J. Fuerst, U.S. Pat. Appl. Pub. No. 2009/093015, published 9 Apr. 2009, Beta-cryptoxanthin production using a novel lycopene beta-monocyclase gene). Hoshino et al. (2006) have disclosed a process for producing zeaxanthin and beta-cryptoxanthin which comprises cultivating a recombinant microorganism expressing beta-carotene hydroxylase gene (Phaffia) under aerobic conditions in aqueous nutrient media and isolating the resulting carotenoids from the cells of the recombinant microorganism or from the broth (T. Hoshino, K. Ojima and Y. Setoguchi, U.S. Pat. Appl. Pub. No. 2006/0121557, published 8 Jun. 2006, Process for producing zeaxanthin and beta-cryptoxanthin).
No microorganism appears able to naturally produce beta-cryptoxanthin as the final product, and hence fermentation technology is not feasible for commercial production. Further, in the fermentation processes the carotenoids produced are low in concentration and as complex mixtures of products, including various added ingredients. Extensive purification steps require large amounts of solvents and the generation of considerable amounts of by-products.
Synthetic Production
Khachik and coworkers developed three processes for the preparation of beta-cryptoxanthin. Two methods employ lutein or lutein esters as the starting material and in the presence of acid it is converted into three forms of anhydroluteins: 3-hydroxy-3′,4′-didehydro-beta-gamma-carotene (I), 3-hydroxy-2′,3′-didehydro-beta-epsilon-carotene (II) and 3-hydroxy-3′,-4′-didehydro-beta-beta-carotene (III). The mixture of anhydroluteins rich in anhydrolutein (III) was subjected to ionic hydrogenation in the presence of an acid and chlorinated solvent to produce alpha- and beta-cryptoxanthin. The purified product showed total carotenoids 85%, of which beta-cryptoxanthin was 55 to 61%, alpha-cryptoxanthin 18 to 30%, and the remainder 3 to 8% R,R-zeaxanthin and un-reacted anhydroluteins (F. Khachik, U.S. Pat. No. 7,115,786 B2, Oct. 3, 2006, Method for production of beta-cryptoxanthin and alpha-cryptoxanthin from commercially available lutein; F. Khachik, A. N. Chang, A. Gana and E. Mazzola, Partial synthesis of (3R,6′R)-alpha-cryptoxanthin and (3R)-beta-cryptoxanthin from (3R,3′R,6′R)-lutein (J. Nat. Products 70, 220-226, 2007)).
In the second method, the mixture of anhydroluteins were converted to alpha- and beta-cryptoxanthin by catalytic hydrogenation using platinum supported on alumina. The final product was reddish crystals with total carotenoids 60% and the HPLC composition showing beta-cryptoxanthin and alpha-cryptoxanthin in the ratio 3:1, 7:3, or 5:1 and the presence of un-reacted anhydroluteins I and II and R,R-zeaxanthin.
In the third method, the process is for the synthesis of optically active 3-hydroxy-beta-ionone and transformation to beta-cryptoxanthin using Wittig coupling reactions. The synthetic approach involves multiple step reactions and purifications leading to a mixture of beta-cryptoxanthin and R,R-zeaxanthin (F. Khachik and A. N. Chang, U.S. Pat. Appl. Pub. No. 2009/0311761, published 17 Dec. 2009, Process for synthesis of 3(S)- and (3R)-3-hydroxy-beta-ionone and their transformation to zeaxanthin and beta-cryptoxanthin; Synthesis of (3S)- and (3R)-hydroxy-beta-ionone and their transformation into (3S)- and (3R)-beta-cryptoxanthin, Synthesis 3, 509-516, 2011).
The Present Invention
As demonstrated by the discussion above, prior processes for producing beta-cryptoxanthin have several limitations. While natural sources of beta-cryptoxanthin are available, extracts have thus far been produced only in enriched form in fruit drinks such as tangerine, Satsuma orange and persimmon. The use of a biotechnological route for producing beta-cryptoxanthin is still in preliminary development and has thus far been limited to laboratory scale production with poor yields. The synthetic approach gives a mixture of beta-cryptoxanthin and a considerable amount of impurities such as alpha-cryptoxanthin, which is most likely zeinoxanthin (a non-provitamin A), along with un-reacted anhydroluteins and zeaxanthin. Applying the processes of Khachik and coworkers, the separation of beta-cryptoxanthin is complex, involves multiple steps and is not commercially feasible. Thus, a need exists for natural beta-cryptoxanthin concentrates of high purity and a process for producing the same.
Based on the chemical structure of anhydrolutein II (3-Hydroxy-2′,3′-didehydro-beta,epsilon-carotene), one would expect hydrogenation at 2′,3′-double bond to form zeinoxanthin (beta,epsilon-caroten-3-ol). In addition, one would expect no conversion of zeinoxanthin to beta-cryptoxanthin by alkali isomerization due to the absence of allylic hydroxyl group. This has been confirmed by phenyl carbinol alkali catalyzed reaction at high temperature (110° C.) of beta-cryptoxanthin containing zeinoxanthin 10% (by HPLC), where after the resultant product showed no change in HPLC profile compared to the control. In fact, the base catalyzed reaction of so called alpha-cryptoxanthin failed to arrive at beta-cryptoxanthin (F. Khachik, A. N. Chang, A. Gana and E. Mazzola, Partial synthesis of 3(R,6′R)-alpha-cryptoxanthin and (3R)-beta-cryptoxanthin from (3R,3R′,6′R)-lutein, J. Nat. Prod. 70, 220-226, 2007).
Review of the art demonstrates the general unavailability of high purity beta-cryptoxanthin, produced in appreciable amounts, as a major ingredient derived from natural sources for use as a nutritional ingredient and in dietary supplements. A primary reason for the unavailability of beta-cryptoxanthin from natural sources (particularly fruits and vegetables) is its low concentration in natural sources, preventing commercialization of this molecule by traditional solvent based extraction procedures. Identifying a source and process to provide commercially available beta-cryptoxanthin concentrates of high purity would help to meet the need for this product and help establish the potential health benefits of beta-cryptoxanthin in clinical trials and as dietary supplements. Capsicum extract has a reasonably high content of beta-cryptoxanthin, but there are many other promising options of beta-cryptoxanthin-containing materials. The present invention meets the need in the art and provides natural beta-cryptoxanthin concentrates of high purity from plant oleoresin, particularly Capsicum oleoresin, and a process for its preparation. In addition, these natural beta-cryptoxanthin concentrates can be used to provide several health benefits, for example in bone loss and osteoporosis.
Bone mass decreases with increasing age. This decrease is due to increased bone resorption and reduced bone formation. The decrease in bone mass induces osteoporosis (M. Yamaguchi, S. Uchiyama, K. Ishiyama, and K. Hashimoto, Oral Administration in Combination with Zinc Enhances Beta-cryptoxanthin-Induced Anabolic Effects on Bone Components in the Femoral Tissues of Rats In Vivo, Biol. Pharm. Bull. 29(2) 371-374 (2006)). Bone homeostasis is maintained through a balance between osteoblastic bone formation and osteoclastic bone resorption (M. Yamaguchi, Role of Carotenoid Beta-Cryptoxanthin in Bone Homeostasis, Journal of Biomedical Science 19-36 (2012)). Production of estrogen decreases in menopause causing an imbalance in metabolism (Citrus unshiu extract. Health Ingredient for prevention of osteoporosis health ingredient for whitening and aesthetic ingredient for cosmetic, Product monograph, Ver.3.0HS by Oryza Oil & Fat Chemical Co Ltd.). Beta-cryptoxanthin has been found to have a potential anabolic effect on bone due to stimulating osteoblastic bone formation and inhibiting osteoclastic bone resorption. Oral administration of beta-cryptoxanthin may have a preventive effect on bone loss with increasing age and on osteoporosis. For example, the role of beta-cryptoxanthin obtained from a Capsicum source in strengthening bone and inhibiting bone resorption is demonstrated in Example 4 below with ovariectomized female wistar rats.